Inhibitors of yeast filamentous growth and method of their manufacture

ABSTRACT

The invention broadly relates to the use of α, β-unsaturated fatty acids to inhibit the filamentous growth of fungi and yeasts and to a method for producing same. In particular the invention relates to the use of optionally substituted C8 to C15 α, β-unsaturated fatty acids or salts, esters or amides thereof for inhibiting or retarding the yeast-to-mycelium transition of organisms having a dimorphic life cycle.

FIELD OF THE INVENTION

The invention relates to the use of α, β-unsaturated fatty acids toinbit the filamentous growth of fungi and yeasts and to a method forproducing same. In particular the invention relates to the use ofoptionally substituted C₈ to C₁₅ fatty acids or salts, esters or amidesthereof for inhibiting or retarding the yeast-to-mycelium transition oforganisms having a dimorphic life cycle.

BACKGROUND OF THE INVENTION

Candida albicans

The dimorphic fungus Candida albicans is one of the most significanthuman fungal pathogens, particularly in immuno-compromised patients.This fungus can infect and colonize in a wide range ofmicro-environments in the body including the blood stream, superficialsites in the mucosa and all of the major internal organs, duringsystemic infections.^(1,2)

A distinguishing feature of C. albicans is the yeast-myceliumdimorphism. The yeast-to-mycelium morphogenic transition is one ofseveral essential virulence attributes in human pathogenesis. Theyeast-to-mycelium molphogenic transition is triggered by varioussignals, including serum, high temperature, neutral pH, nutrient poormedia, and certain chemicals such as N-acetylglucosamine. It appearsthat many of the responses to these signals reflect normal interactionsbetween the fungus and its host in vivo.

The yeast-to-mycelium transition has been shown to be one of severalvirulence attributes that enable C. albicans to invade human tissues.³⁻⁵Patients with serious disease generally have filaments of C. albicanspenetrating the infected tissue. Various genes involved in hyphalmorphogenesis have been identified, including cek1, cla4, cpp1, cst20,and tup1, encoding MAP kinase, PAK, phosphates, MEKK kinase, andtranscription factor, respectively. Mutation of these genes blocked C.albicans yeast-to-mycelium transition and attenuated its virulenceagainst mouse animal models.^(6,7)

Several categories of natural and synthetic chemicals have beenevaluated in recent years against C. albicans cell growth and dimorphismtransition. A novel cyclic β-amino acid that inhibits isoleucyl-tRNAsynthetase was shown to inhibit cell growth of C. albicans. ⁸ Carvoneand perillaldehyde were found to interfere with the formation offilamentous structures of C. albicans. ⁹

An autoregulatory substance (ARS) from the eukaryote C. albicans,characterized as 3,7,11-trimethyl-2,6,10-dodecatrienoic acid (farnesoicacid), inhibits C. albicans germ tube (mycelium) formation and appearsto play a key role in the regulation of the morphological transition inC. albicans. ¹⁰ Its derivatives have been evaluated for their activityin regulation of morphological transitions in C. albicans and it wasconcluded that the trans isomer of farnesoic acid is essential for itspotent inhibition of the yeast-to-mycelium transition.¹¹

Farnesol, a close derivative of ARS, was shown to prevent mycelialdevelopment of C. albicans in both growth morphology assay anddifferentiation assay using three chemically distinct activators forgerm tube formation (L-proline, N-acetylglucosamine, and serum).¹²

Farnesol and its metabolite farnesoic acid (ARS) is also found in higherlife forms, including humans. They are generated intracellularly andsome of the farnesyl derivatives are required for synthesis ofcholesterol, steroids, retinoids, and farnesylated proteins.^(16,17)Moreover, farnesol metabolites have been implicated as signallingmolecules in the regulation of cholesterol degradation throughactivation of FXR, a nuclear receptor which represses cholesterolmetabolism pathway.^(16,18) Conceivably, tipping over the subtle balanceof these important intermediates and cellular signals may lead tosubstantial changes in normal physiological processes.

A diffusible signal factor (DSF) from the plant pathogen Xanthomonascampestris is a prokaryotic cell-cell communication signal and requiredfor bacterial virulence,¹³ but little is known about its structure andscope of function.

Rapid emergence of antibiotic resistance demands development ofalternative approaches to prevent and control infectious diseases. Asthe lethal effect of antibiotics is the common cause to force microbesto mutate and survive, non-antimicrobial means of controlling C.albicans and other microorganisms could be useful to avoid or delay thedevelopment of antibiotic-resistance. Inhibiting the yeast-to-myceliumtransition avoids selection of resistant organisms.

Synthesis of α,β-unsaturated Fatty Acids

Methods for producing short chain (less than 8 carbons)cis-α,β-unsaturated fatty acids using the Favorsky rearrangement ofcorresponding 1,3-dibromo-2-ones in alkaline solution at roomtemperature are reported in the art which also reports that best yieldsare obtained when using potassium bicarbonate.^(14,15)

It is an object of the present invention to provide an alternative meansfor controlling Candida albicans infection or to at least provide thepublic with a useful choice.

SUMMARY OF THE INVENTION

According to a first aspect of the present invention there is provideduse of at least one optionally substituted C₈ to C₁₅ fatty acid or asalt, ester or amide thereof containing only one double bond wherein thedouble bond is at the α,β-position in the manufacture of a formulationfor inhibiting or retarding the yeast-to-mycelium transition of anorganism having a yeast-to-mycelium dimorphic life cycle.

A second aspect of the invention provides a method of inhibiting orretarding the yeast-to-mycelium transition of an organism having ayeast-to-mycelium dimomphic life cycle comprising administering to asubject at least one optionally substituted C₈ to C₁₅ fatty acid or asalt, ester or amide thereof containing only one double bond wherein thedouble bond is at the α,β-position.

A third aspect of the invention provides a formulation for inhibiting orretarding the yeast-to-mycelium transition of an organism having ayeast-to-mycelium dimorphic life cycle comprising at least oneoptionally substituted C₈ to C₁₅ fatty acid or a salt, ester or amidethereof containing only one double bond wherein the double bond is atthe α,β-position.

A fourth aspect of the present invention provides use of at least oneoptionally substituted C₈ to C₁₅ fatty acid or a salt, ester or amidethereof containing only one double bond wherein the double bond is atthe α,β-position in the manufacture of a formulation for treating orpreventing an injection by an organism having a yeast-to-myceliumdimorphic life cycle.

A fifth aspect of the invention provides a method treating or preventingan infection by an organism having a yeast-to-mycelium dimorphic lifecycle comprising administering to a subject at least one optionallysubstituted C₈ to C₁₅ fatty acid or a salt, ester or amide thereofcontaining only one double bond wherein the double bond is at theα,β-position.

A sixth aspect of the invention provides a formulation treating orpreventing an infection by an organism having a yeast-to-myceliumdimorphic life cycle comprising at least one optionally substituted C₈to C₁₅ fatty acid or a salt, ester or amide thereof containing only onedouble bond wherein the double bond is at the α,β-position.

The following statements relate to the above aspects of the invention.

In one embodiment, the organism having a yeast-to-mycelium dimorphiclife cycle is selected from the group comprising Aspergillus fumigatus,Aureobasidium pullulans, Benjaminiella poitrasii, Blastomycesdermatitidis, Cerato cystis ulmi, Debaryomyces hansenii, Histoplasmacapsulatum, Paracoccidioides brasiliensis, Phaeococcomyces exophiale,Sporothrix schenckii, Ustilago maydis, Yarrowia lipolytica and organismsof the Candida genus. A preferred organism of the Candida genus isCandida albicans.

In one embodiment, preferably the fatty acid is a C₈ to C₁₃ fatty acid.Preferably the fatty acid is a C₁₂ fatty acid.

In another embodiment, preferably the fatty acid has at least onesubstituent. In one embodiment the substituent is a branched alkylgroup. Preferably the substituent is an alkyl group, preferably a groupselected from the group comprising methyl, ethyl, propyl, butyl, andpentyl, and isomeric forms thereof. It will be apparent to a skilledworker that the fatty acid may have other substituents.

In another embodiment, preferably the fatty acid is selected from thegroup consisting of:

(CH₃)₂CH(CH₂)₇CH═CHCOOH;

CH₃(CH₂)₄CH═CHCOOH;

CH₃(CH₂)₆CH═CHCOOH;

CH₃(CH₂)₈CH═CHCOOH;

CH₃(CH₂)₉CH═CHCOOH; and

CH₃(CH₂)₁₁CH═CHCOOH.

In another embodiment, preferably the fatty acid is a trans fatty acid.

In another embodiment, preferably the fatty acid is a cis fatty acid.

In another embodiment, preferably the formulation comprises a food,drink, food additive, drink additive, dietary supplement, nutraceuticalor pharmaceutical composition. Preferably the pharmaceutical compositioncomprises a fatty acid of the invention and a pharmaceuticallyacceptable carrier.

A seventh aspect of the present invention provides a process forproducing a cis α,β-unsaturated fatty acid of the formula R—CH═CH—COOHcomprising reacting a compound of Formula I:

with:

(1) NaOH; followed by

(2) an acid,

wherein R is an optionally substituted alkyl group and X and Y areindependently selected from the group comprising Cl and Br.

In one embodiment, preferably R is optionally substitutedCH₃(CH₂)₃₋₁₀CH₂— or (CH₃)₂CH(CH₂)₂₋₉CH₂—.

In one embodiment, preferably R is has at least one substituent. In apreferred embodiment the substituent is a branched alkyl group.Preferably the substituent is an alkyl group, preferably a groupselected from the group comprising methyl, ethyl, propyl, butyl, andpentyl, and isomeric forms thereof. It will be apparent to a skilledworker that R may have other substituents.

In one embodiment, preferably a compound of formula (I) is reacted withabout 0.5, 1 2, 3, 4, 5 or 6 equivalent NaOH.

In one embodiment, preferably a compound of formula (I) is reacted withNaOH for about 0.5, 1, 2, 3,4, 5, 6, 7, 8, or 9 hours or longer.

In one embodiment, preferably a compound of formula (I) is reacted withNaOH for about 4 to about 8 hours.

In one embodiment, preferably the acid comprises about 0.5, 1, 1.5 or 2N HCl.

In one embodiment, preferably a compound of formula (I) is reacted with4 equivalent NaOH and H₂O for about 6 to about 8 hours followed byneutralization with 1N HCl.

In one embodiment, preferably the process is carried out at about 20 to30 degrees C.

An eighth aspect of the invention provides a cis α,β-unsaturated fattyacid produced by a process of the invention.

A ninth aspect of the invention provides cis-11-methyl-2-dodecenoicacid.

A tenth aspect of the invention provides trans-11-methyl-2-dodecenoicacid.

An eleventh aspect of the invention provides a pharmaceuticalcomposition comprising cis and/or trans 11-methyl-2-dodecenoic acid anda pharmaceutically acceptable carrier.

A twelfth aspect of the invention provides a pharmaceutical compositioncomprising a cis and/or trans fatty acid selected from the groupcomprising CH₃(CH₂)₄CH═CHCOOH; CH₃(CH₂)₆CH═CHCOOH; CH₃(CH₂)₈CH═CHCOOH;CH₃(CH₂)₉CH═CHCOOH; and CH₃(CH₂)₁₁CH═CHCOOH and a pharmaceuticallyacceptable carrier.

This invention may also be said broadly to consist in the part, elementsand features referred to or indicated in the specification of theapplication, individually or collectively, and any or combinations ofany two or more said parts, elements or features, and where specificallyintegers are mentioned herein which have known equivalents in the art towhich this invention relates, such known equivalents are deemed to beincorporated herein as if individually set forth.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 a shows HPLC analysis of the efficiency of synthesisingcis-2-dodecenoic acid using a previously reported method.

FIG. 1 b shows HPLC analysis of the efficiency of synthesisingcis-2-dodecenoic acid using a method of the invention.

FIG. 2 shows the in vitro inhibitory activity of selected fatty acidsagainst the yeast-to-mycelium transition of Candida albicans.

FIGS. 3 a and 3 b show that DSF does not affect yeast growth butinhibits the yeast-to-mycelium transition of a culture of Candidaalbicans (the bar indicates 30 μm).

DETAILED DESCRIPTION OF THE INVENTION

The applicants have found that α,β-unsaturated fatty acids that arestructurally dissimilar to ARS have efficacy in inhibiting or retardingthe yeast-to-mycelium transition of organisms having a yeast-to-myceliumdimorphic life cycle and that cis isomers also have efficacy contrary toprevious reports. Exemplary organisms having a yeast-to-myceliumdimorphic life cycle include organisms of the Candida genus such asCandida albicans. The invention is also useful in respect of otherorganisms having a yeast-to-mycelium dimorphic life cycle including butnot limited to Aspergillus fumigatus, Aureobasidium pullulans,Benjaminiella poitrasii, Blastomyces dermatitidis, Ceratocystis ulmi,Debaryomyces hansenii, Histoplasma capsulatum, Paracoccidioidesbrasiliensis, Phaeococcomyces exophiale, Sporothrix schenckii, Ustilagomaydis, and Yarrowia lipolytica. The fatty acids of the invention arethus useful to treat or prevent infection by these organisms.

Fatty acids useful herein include optionally substituted C₈ to C₁₅α,β-unsaturated fatty acids or salts, esters or amides thereofcontaining one double bond wherein the double bond is at theα,β-position. In one embodiment, preferably the fatty acid is a C₈ toC₁₃ fatty acid. Preferably the fatty acid is a C₁₂ fatty acid. Fattyacids useful herein may be substituted or unsubstituted, and cis ortrans fatty acids. Fatty acids useful herein include(CH₃)₂CH(CH₂)₇CH═CHCOOH; CH₃(CH₂)₄CH═CHCOOH; CH₃(CH₂)₆CH═CHCOOH;CH₃(CH₂)₈CH═CHCOOH; CH₃(CH₂)₉CH═CHCOOH; and CH₃(CH₂)₁₁CH═CHCOOH. It willbe apparent to a skilled worker that the fatty acid may comprise alkylor other substituents.

The applicants have also elucidated the structure of DSF to becis-11-methyl-2-dodecenoic acid.

The applicants have also elucidated a novel reaction scheme for highyield production of cis-α,β-unsaturated fatty acids.

The previously reported methods for synthesis of α,β-unsaturated fattyacids can be used for preparation of short-chain cis-α,β-unsaturatedfatty acids (less than 8 carbons).⁵

However, it was found that these methods were not suitable forsynthesising long-chain cis-α,β-unsaturated fatty acids. For example,under the reported optimal reaction conditions, the yield ofcis-2-dodecenoic acid (FIG. 2, compound 7) was found to be less than 5%,even after a prolonged reaction time of up to 96 h (FIG. 1 a-first peakcis-2-dodecenoic acid; second peak 1,3-dibromo-2-dodecanone).

It was found that replacing potassium bicarbonate with sodium hydroxideincreased the yield of cis-2-dodecenoic acid up to 98% (Scheme 1 andFIG. 1 b-first peak cis-2-dodecenoic acid; second peak1,3-dibromo-2-dodecanone).

In Scheme 1, (a) LiMe, CH₃OCH₃; (b) H₂O; (c) Br₂, HBr, H₂O; (d) NaOH,H₂O; (e) HCI.

In preferred embodiment of Scheme 1, (a) LiMe, CH₃OCH₃, 4 h; (b) H₂O,˜90% yield; (c) Br₂, HBr, H₂O, 2 h: ˜50% yield; (d) NaOH, H₂O, 6-8 h;(e) 1N HCI, >85% yield.

Accordingly, in one aspect, the invention provides a process forproducing a cis α,β-unsaturated fatty acid of the formula R—CH═CH—COOHcomprising reacting a compound of Formula I:

with:

(1) NaOH; followed by

(2) an acid,

wherein R is an optionally substituted alkyl group and X and Y areindependently selected from the group comprising Cl and Br.

In one embodiment, preferably R is optionally substitutedCH₃(CH₂)₃₋₁₀CH₂— or (CH₃)₂CH(CH₂)₂₋₉CH₂—.

In one embodiment, preferably R is substituted. In a preferredembodiment the substitution is a branched alkyl group. Preferably thesubstituted is an alkyl group, preferably a group selected from thegroup comprising methyl, ethyl, propyl, butyl, and pentyl, and isomericforms thereof. It will be apparent to a skilled worker that R may haveother substituents.

In one embodiment, preferably a compound of formula (I) is reacted withabout 0.5, 1 2, 3,4, 5 or 6 equivalent NaOH.

In one embodiment, preferably a compound of formula (I) is reacted withNaOH for about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, or 9 hours or longer.

In one embodiment, preferably a compound of formula (I) is reacted withNaOH for about 4 to about 8 hours.

In one embodiment, preferably the acid comprises about 0.5, 1, 1.5 or 2equivalent HCl.

In one embodiment, preferably a compound of formula (I) is reacted with4 equivalent NaOH and H₂O for about 6 to about 8 hours followed byneutralization with 1N HCl.

In one embodiment, preferably the process is carried out at about 20 to30 degrees C.

To conduct the HPLC analyses depicted in FIGS. 1 a and 1 b, the reactionproducts were extracted with hexane, dried, then analyzed using HPLC ona C₁₈ reverse-phase column (4.6×250 mm) and eluted with methanol-water(80:20, vol/vol) at a flow rate of 1 ml/min. The retention time was 9.5min for cis-2-dodecenoic acid, and 23.2 min for1,3-dibromo-2-dodecanone, respectively.

The other long-chain cis-α,β-unsaturated fatty acids of FIG. 2 were alsosynthesized in high yields using the modified method of Scheme 1 (Table1). TABLE 1 Yields of cis-α,β-unsaturated fatty acids produced accordingto Scheme 1 Compound* 1 5 6 7 8 9 Yield (%) 88.3 93.9 96.5 98.1 97.295.5*Compounds are numbered according to FIG. 2. The yield was calculated asa percentage of product/substrate based on HPLC analysis.

The purified reaction products were examined by NMR. In all cases, thechemical shifts and the coupling constants of the protons on the doublebond (˜11.5 Hz) indicate the products were cis-acids (data not shown).

FIG. 2 shows the in vitro inhibitory activity of selected fatty acidsagainst the yeast-to-mycelium transition of Candida albicans, includingthe cis and trans isomers of DSF (compounds 1 and 2), ARS (compound 3),and compounds produced according to a process of the invention (ciscompounds 5 to 9).

Contrary to the previous reports that the trans configuration of theα,β-double bond of ARS is essential for the inhibitory effect against C.albicans filamentous growth,¹¹ DSF and compound 2 (the trans-isomer ofDSF) showed an identical inhibitory activity. Moreover, the other cis-and trans-isomer pairs of Ε,β-unsaturated fatty acids (compounds 5 and11, 6 and 12, 7 and 13, 8 and 14, 9 and 15) also each showed verysimilar or identical inhibitory activity (FIG. 2).

As yeast-to-mycelium transition appears to be necessary for thepathogenicity of C. albicans, the compounds disclosed herein haveutility for the control of the fungal disease. Rapid emergence ofantibiotic resistance demands development of alternative approaches toprevent and control infectious diseases. As the lethal effect ofantibiotics is the common cause to force microbes to mutate and survive,non-antimicrobial means of controlling C. albicans and othermicroorganisms could be useful to avoid or delay the development ofantibiotic-resistance. Inhibiting the yeast-to-mycelium transitionprovides such a possibility.

The fatty acids useful herein may be incorporated into a food, drink,food additive, drink additive, dietary supplement, neutraceutical orpharmaceutical.

The pharmaceutical compositions provided by the invention comprise afatty acid of the invention and a pharmaceutically acceptable carrier.The pharmaceutical compositions of the invention can be administered inany suitable manner and thus may be formulated for administrationorally, topically, subcutaneously, intramuscularly, intravenously, orparenterally, for example.

The invention consists in the foregoing and also envisages constructionsof which the following gives examples only.

EXAMPLE 1

Scheme 1 and Scheme 2 set out below were employed to synthesizeα,β-unsaturated fatty acids listed in FIG. 2. As shown in Scheme 1,cis-α,β-unsaturated fatty acids were synthesized using the Favorskyrearrangement of corresponding 1,3-dibromo-2-ones in alkaline solutionat room temperature by modifying a previously reported method.^(15,16)Commercially available fatty acids (Sigma-Aldrich, USA) were convertedto corresponding methyl ketones in the presence of LiMe. Methyl ketoneswere reacted with bromide reagents to produce 1,3-dibromo-2-ones whichwere then rearranged to produce cis-α,β-unsaturated fatty acids in thepresence of 4 equivalent NaOH.

In Scheme 1, (a) LiMe, CH₃OCH₃, 4 h; (b) H₂O, ˜90% yield; (c) Br₂, HBr,H₂O, 2 h, ˜50% yield; (d) NaOH, H₂O, 6-8 h; (e) 1N HCI, >85% yield.

Trans-α,β-unsaturated fatty acids were synthesized through eliminationof 2-bromo fatty acids prepared by brominating saturated fatty acids,using potassium tert-butoxide as a base in tert-butanol (Scheme 2). Themethyl ester of ARS (3,7,11-trimethyl-2,6,10-dodecatrienoic acid) wassynthesized as previously reported¹⁶ and then hydrolyzed to produce ARS.

In Scheme 2, (a) Br₂, HBr, H₂O, 2 h, ˜55% yield; (b) KOBu^(tert),tert-BuOH, 3 h; (c) 1N HCl, >90% yield.

Referring to FIG. 2, compounds 1 and 5-9 were synthesized according toScheme 1, compounds 2 and 11-15 were synthesized according to Scheme 2,compound 3 was synthesized as previously reported¹⁶, and the others wereobtained commercially from Sigma-Aldrich, USA. All the syntheticcompounds were characterized by ¹H NMR, ¹³C NMR, and mass spectroscopy.

EXAMPLE 2

An in vitro bioassay of the inhibitory activity of the fatty acids ofFIG. 2 against C. albicans yeast-to-mycelium transition was carried out.

C. albicans strain CAI4 (ATCC MYA-684) was incubated at 28° C. for 24 hwith shaking in a glucose salts (GS) medium (5 g of glucose, 0.26 g ofNa₂HPO₄.12H₂O, 0.66 g of KH₂PO₄, 0.88 g of MgSO₄.7H₂O, 0.33 g of NH₄Cl,16 μg of biotin per litre) supplemented with 0.1 mg/ml of uridine.

All the C. albicans cells remained, in the yeast form under theseconditions. The yeast cells were washed three times with GS mediumbefore use and then were dispersed in GS medium to give a solution witha final OD₆₀₀ of 1.0.

The test compounds set out in FIG. 2 were diluted to the indicatedconcentrations with GS medium and were added to 100 μl of the washed C.albicans culture in 96-well microtitre plates, mixed and incubated at37° C. for 6 hours (h).

The percentages of yeast cells and germ tubes (filamentous form) weredetermined microscopically by counting about 300 cells from theereplicates. Inhibitory activity was defined as the mole concentrationthat gave a minimum of 90% inhibition (IC₉₀) relative to control.

Compounds with useful yeast-to-mycelium inhibitory activity werecompounds 1, 2, 5 to 8 and 11 to 14 as shown in FIG. 2.

EXAMPLE 3

Compound 1 was not observed to have a toxic effect on yeast cell growtheven at concentrations up to 1 mM, as shown in FIG. 3 b (the barindicates 30 μm).

A starter culture of C. albicans strain CA14 was prepared by growing thefungus at a low temperature (28° C.) in GS medium to maintain the yeastcell morphology. The culture was diluted with the same fresh medium withor without supplement of DSF and incubated at a high temperature (37°C.) which promotes yeast-to-mycelium conversion. FIG. 3 a (the barindicates 30 μm) shows that within 6 hours of incubation, the majorityof C. albicans cells formed germ tubes in the absence of DSF, whereasaddition of 30 μM DSF to the culture of C. albicans completely abolishedyeast to-mycelium transition (FIG. 3 b). No toxic effect on yeast cellgrowth was noticed even when DSF concentration was increased up to 1 mM.The synthetic DSF was indistinguishable with the natural DSF ininhibition of germ tube formation.

The above describes some preferred embodiments of the present inventionand indicates several possible modifications but it will be appreciatedby those skilled in the art that other modifications can be made withoutdeparting from the scope of the invention.

REFERENCES

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1. Use of at least one optionally substituted C₈ to C₁₅ fatty acid or asalt, ester or amide thereof containing only one double bond wherein thedouble bond is at the α,β-position in the manufacture of a formulationfor inhibiting or retarding the yeast-to-mycelium transition of anorganism having a yeast-to-mycelium dimorphic life cycle.
 2. Use of atleast one optionally substituted C₈ to C₁₅ fatty acid or a salt, esteror amide thereof containing only one double bond wherein the double bondis at the α,β-position in the manufacture of a formulation for treatingor preventing an infection by an organism having a yeast-to-myceliumdimorphic life cycle.
 3. A use of claim 1 or 2 wherein the organism isselected from the group comprising Aspergillus fumigatus, Aureobasidiumpullulans, Benjaminiella poitrasii, Blastomyces dermatitidis,Ceratocystis pullulans, Benjaminiella Histoplasma capsulatum,Paracoccidioides brasiliensis, Phaeococcomyces exophiale, Sporothrixschenckii, Ustilago maydis, Yarrowia lipolytica and organisms of theCandida genus.
 4. A use of claim 1 or 2 wherein the organism is Candidaalbicans.
 5. A use of any one of claims 1 to 4 wherein the fatty acid isa C₈ to C₁₃ fatty acid.
 6. A use of any one of claims 1 to 4 wherein thefatty acid is a C₁₂ fatty acid.
 7. A use of any one of the precedingclaims wherein the fatty acid is substituted.
 8. A use of claim 7wherein the substituent comprises a branched alkyl group.
 9. A use ofclaim 7 wherein the substituent comprises an alkyl group selected fromthe group comprising methyl, ethyl, propyl, butyl, and pentyl, andisometric forms thereof.
 10. A use of any one of claims 1 to 4 whereinthe fatty acid is selected from the group consisting of:(CH₃)₂CH(CH₂)₇CH═CHCOOH; CH₃(CH₂)₄CH═CHCOOH; CH₃(CH₂)₆CH═CHCOOH;CH₃(CH₂)₈CH═CHCOOH; CH₃(CH₂)₉CH═CHCOOH; and CH₃(CH₂)₁₁CH═CHCOOH.
 11. Ause of any one of claims 1 to 10 wherein the fatty acid is a trans fattyacid.
 12. A use of any one of claims 1 to 10 wherein the fatty acid is acis fatty acid.
 13. A use of any one of the preceding claims wherein theformulation comprises a food, drink, food additive, drink additive,dietary supplement, nutraceutical or pharmaceutical.
 14. A method ofinhibiting or retarding the yeast-to-mycelium transition of an organismhaving a yeast-to-mycelium dimorphic life cycle comprising administeringto a subject at least one optionally substituted C₈ to C₁₅ fatty acid ora salt, ester or amide thereof containing only one double bond whereinthe double bond is at the α,β-position.
 15. A method treating orpreventing an infection by an organism having a yeast-to-myceliumdimorphic life cycle comprising administering to a subject at least oneoptionally substituted C₈ to C₁₅ fatty acid or a salt, ester or amidethereof containing only one double bond wherein the double bond is atthe α,β-position.
 16. A method of claim 14 or 15 wherein the organism isselected from the group comprising Aspergillus fumigatus, Aureobasidiumpullulans, Benjaminiella poitrasii, Blastomyces dermatitidis,Ceratocystis ulmi, Debaryomyces hansenii, Histoplasma capsulatum,Paracoccidioides brasiliensis, Phaeococcomyces exophiale, Sporothrixschenckii, Ustilago maydis, Yarrowia lipolytica and organisms of theCandida genus.
 17. A method of claim 14 or 15 wherein the organism isCandida albicans.
 18. A method of any one of claims 14 to 17 wherein thefatty acid is a C₈ to C₁₃ fatty acid.
 19. A method of any one of claims14 to 17 wherein the fatty acid is a C₁₂ fatty acid.
 20. A method of anyone of claims 14 to 19 wherein the fatty acid is substituted.
 21. Amethod of claim 20 wherein the substituent comprises a branched alkylcroup.
 22. A method of claim 20 wherein the substituent comprises analkyl group selected from the group comprising methyl, ethyl, propyl,butyl, and pentyl, and isomeric forms thereof.
 23. A method of any oneof claims 14 to 17 wherein the fatty acid is selected from the croupconsisting of: (CH₃)₂CH(CH₂)₇CH═CHCOOH; CH₃(CH₂)₄CH═CHCOOH;CH₃(CH₂)₆CH═CHCOOH; CH₃(CH₂)₈CH═CHCOOH; CH₃(CH₂)₉CH═CHCOOH; andCH₃(CH₂)₁₁CH═CHCOOH.
 24. A method of any one of claims 14 to 23 whereinthe fatty acid is a trans fatty acid.
 25. A method of any one of claims14 to 23 wherein the fatty acid is a cis fatty acid.
 26. A formulationfor inhibiting or retarding the yeast-to-mycelium transition of anorganism having a yeast-to-mycelium dimorphic life cycle comprising atleast one optionally substituted C₈ to C₁₅ fatty acid or a salt, esteror amide thereof containing only one double bond wherein the double bondis at the α,β-position.
 27. A formulation treating or preventing aninfection by an organism having a yeast-to-mycelium dimorphic life cyclecomprising at least one optionally substituted C₈ to C₁₅ fatty acid or asalt, ester or amide thereof containing only one double bond wherein thedouble bond is at the α,β-position.
 28. A formulation of claim 26 or 27wherein the organism is selected from the group comprising Aspergillusfumigatus, Aureobasidium pullulans, Benjaminiella poitrasii, Blastomycesdermatitidis, Ceratocystis ulmi, Debaryomyces hansenii, Histoplasmacapsulatum, Paracoccidioides brasiliensis, Phaeococcomyces exophiale,Sporothrix schenckii, Ustilago maydis, Yarrowia lipolytica and organismsof the Candida genus.
 29. A formulation of claim 26 or 27 wherein theorganism is Candida albicans.
 30. A formulation of any one of claims 26to 29 wherein the fatty acid is a C₈ to C₁₃ fatty acid.
 31. Aformulation of any one of claims 26 to 29 wherein the fatty acid is aC₁₂ fatty acid.
 32. A formulation of any one of claims 26 to 31 whereinthe fatty acid is substituted.
 33. A formulation of claim 32 wherein thesubstituent comprises a branched alkyl group.
 34. A formulation of claim32 wherein the substituent comprises an alkyl group selected from thegroup comprising methyl, ethyl, propyl, butyl, and pentyl, and isomericforms thereof.
 35. A formulation of any one of claims 26 to 29 whereinthe fatty acid is selected from the group consisting of:(CH₃)₂CH(CH₂)₇CH═CHCOOH; CH₃(CH₂)₄CH═CHCOOH; CH₃(CH₂)₆CH═CHCOOH;CH₃(CH₂)₈CH═CHCOOH; CH₃(CH₂)₉CH═CHCOOH; and CH₃(CH₂)₁₁CH═CHCOOH.
 36. Aformulation of any one of claims 26 to 35 wherein the fatty acid is atrans fatty acid.
 37. A formulation of any one of claims 26 to 35wherein the fatty acid is a cis fatty acid.
 38. A formulation of any oneof claims 26 to 37 which comprises a food, drink, food additive, drinkadditive, dietary supplement, nutraceutical or pharmaceuticalcomposition.
 39. A process for producing a cis α,β-unsaturated fattyacid of the formula R—CH═CH—COOH comprising reacting a compound ofFormula I:

with: (1) NaOH; followed by (2) an acid, wherein R is an optionallysubstituted alkyl group and X and Y are independently selected from thegroup comprising Cl and Br.
 40. A process of claim 39 wherein R isoptionally substituted CH₃(CH₂)₃₋₁₀CH₂— or (CH₃)₂CH(CH₂)₂₋₉CH₂—.
 41. Aprocess of claim 39 or 40 wherein R is substituted.
 42. A process ofclaim 41 wherein the substituent comprises a branched alkyl group.
 43. Aprocess of claim 41 wherein the substituent comprises an alkyl groupselected from the group comprising methyl, ethyl, propyl, butyl, andpentyl, and isomeric forms thereof.
 44. A process of any one of claims39 to 43 wherein a compound of formula (I) is reacted with about 0.5, 1,2, 3, 4, 5 or 6 equivalent NaOH.
 45. A process of any one of claims 39to 44 wherein a compound of formula (I) is reacted with NaOH for about0.5, 1, 2, 3, 4, 5, 6, 7, 8, or 9 hours.
 46. A process of any one ofclaims 39 to 43 wherein a compound of formula (I) is reacted with NaOHfor about 6 to about 8 hours.
 47. A process of anyone of claims 39 to 46wherein the acid comprises about 0.5, 1, 1.5 or 2 N HCl.
 48. A processof any one of claims 39 to 43 wherein a compound of formula (I) isreacted with 4 equivalent NaOH and H₂O for about 4 to about 8 hoursfollowed by 1N HCl.
 49. A process of any one of claims 39 to 48 carriedout at about 20 to 30 degrees C.
 50. A cis α,β-unsaturated fatty acidwhen produced by a process as claimed in any one of claims 39 to
 49. 51.cis or trans 11-methyl-2-dodecenoic acid.
 52. A pharmaceuticalcomposition comprising cis or trans 11-methyl-2-dodecenoic acid and apharmaceutically acceptable carrier.
 53. A pharmaceutical compositioncomprising a cis or traps fatty acid selected from the group comprising:CH₃(CH₂)₄CH═CHCOOH; CH₃(CH₂)₆CH═CHCOOH; CH₃(CH₂)₈CH═CHCOOH;CH₃(CH₂)₉CH═CHCOOH; and CH₃(CH₂)₁₁CH═CHCOOH and a pharmaceuticallyacceptable carrier.